A Comparative Study on the Recovery of  EcoRI Endonuclease from Two Different Genetically Modified Strains of Escherichia coli

Candan TAMERLER; Z. İlsen ÖNSAN; Betül KIRDAR

Makale detayları

A Comparative Study on the Recovery of EcoRI Endonuclease from Two Different Genetically Modified Strains of Escherichia coli

Yazarlar:

Candan TAMERLER | Z. İlsen ÖNSAN | Betül KIRDAR

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Chemistry 2001 Vol.25 Issue 1, pp.63-71

Belge Tipi:

Makale

Yayın Tarihi:

12/2000

Belge Dili:

Türkçe

Document ID (Org No.):

2000730182

Sıklık:

İki Aylık

eISSN:

1303-6130

pISSN:

1300-0527

Özet:

 

A laboratory scale procedure developed for the purification of   EcoRI restriction endonuclease was applied to two different   Escherichia coli} strains,  E. coli 294 and   E. coli M5248, which are genetically modified to  overproduce the enzyme.  The purification method consisted of three  successive chromatographic steps including phosphocellulose and  hydroxyapatite columns and further fractionation in a second  phosphocellulose column.  It was shown that the second  phosphocellulose separation can be omitted in the case of  E.  coli 294.  Quality control tests indicated enzyme preparations free  of contaminants and endo- or exo-nucleases.  The yields obtained at  the final stage of the purification were 1.3x105 U/g cells for   E. coli M5248 and 3.3x106 U/g cells for  E.  coli 294.

 

A laboratory scale procedure developed for the purification of   EcoRI restriction endonuclease was applied to two different   Escherichia coli} strains,  E. coli 294 and   E. coli M5248, which are genetically modified to  overproduce the enzyme.  The purification method consisted of three  successive chromatographic steps including phosphocellulose and  hydroxyapatite columns and further fractionation in a second  phosphocellulose column.  It was shown that the second  phosphocellulose separation can be omitted in the case of  E.  coli 294.  Quality control tests indicated enzyme preparations free  of contaminants and endo- or exo-nucleases.  The yields obtained at  the final stage of the purification were 1.3x105 U/g cells for   E. coli M5248 and 3.3x106 U/g cells for  E.  coli 294.

Makale detayları

A Comparative Study on the Recovery of EcoRI Endonuclease from Two Different Genetically Modified Strains of Escherichia coli

Yazarlar: Candan TAMERLER | Z. İlsen ÖNSAN | Betül KIRDAR

Yayıncı: TUBITAK

Dergi: Turkish Journal of Chemistry 2001 Vol.25 Issue 1, pp.63-71

Belge Tipi: Makale

Yayın Tarihi: 12/2000

Belge Dili: Türkçe

Document ID (Org No.): 2000730182

Sıklık: İki Aylık

eISSN: 1303-6130

pISSN: 1300-0527

Sayı

Yazarlar:

Candan TAMERLER | Z. İlsen ÖNSAN | Betül KIRDAR

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Chemistry 2001 Vol.25 Issue 1, pp.63-71

Belge Tipi:

Makale

Yayın Tarihi:

12/2000

Belge Dili:

Türkçe

Document ID (Org No.):

2000730182

Sıklık:

İki Aylık

eISSN:

1303-6130

pISSN:

1300-0527